Lymphotoxin-alpha/TNF-beta Antibody (3F12.2D3) - Chimeric - Azide and BSA Free
Novus Biologicals, part of Bio-Techne | Catalog # NBP2-81105
Recombinant Monoclonal Antibody
![Western Blot: Lymphotoxin-alpha/TNF-beta Antibody (3F12.2D3)ChimericAzide and BSA Free [NBP2-81105]](https://resources.bio-techne.com/images/products/Lymphotoxin-alpha-TNF-beta-Antibody-3F12-2D3-Chimeric-Western-Blot-NBP2-81105-img0002.jpg "Western Blot: Lymphotoxin-alpha/TNF-beta Antibody (3F12.2D3)ChimericAzide and BSA Free [NBP2-81105]")
Conjugate
Catalog #
Key Product Details
Validated by
Biological Validation
Species Reactivity
Validated:
Human
Applications
ELISA, Flow Cytometry, Neutralization, Surface Plasmon Resonance, Western Blot
Label
Unconjugated
Antibody Source
Recombinant Monoclonal Rabbit IgG Kappa Clone # 3F12.2D3
Format
Azide and BSA Free
Concentration
1 mg/ml
Product Summary
Immunogen
3F12 was prepared by hyperimmunizing BALB/c mice with purified recombinant human LTalpha expressed in E. coli - the hybridoma clone was selected for its ability to bind LTalpha3 by ELISA. The CDRs of the antibody were grafted to the framework regions of the mice mAb 2G7 to create the chimeric version of the antibody.
Specificity
3F12 binds to human LTalpha3 and LT-alpha-beta (with a KD of ~0.3 nM, determined by BIACORE; ~ 37 pM, determined by ELISA) in both its soluble homotrimeric and membrane heterotrimeric forms. LTalpha belongs to the tumor necrosis factor (TNF) superfamily and is secreted as a homo-trimer (LTalpha3), or is expressed on the cell surface in complex with LTbeta. Lymphotoxin is produced by lymphocytes and in its homotrimeric form binds to TNFRSF1A/TNFR1, TNFRSF1B/TNFBR and TNFRSF14/HVEM. LTalpha3 signaling induces target cells to upregulate many chemokines and cytokines in an NFkB-dependent manner.
Clonality
Monoclonal
Host
Rabbit
Isotype
IgG Kappa
Scientific Data Images
Western Blot: Lymphotoxin-alpha/TNF-beta Antibody (3F12.2D3)ChimericAzide and BSA Free [NBP2-81105]
Western Blot: Lymphotoxin-alpha/TNF-beta Antibody (3F12.2D3) - Chimeric [NBP2-81105] - Western Blot using Lymphotoxin-alpha/TNF-beta Antibody (3F12.2D3) [NBP2-81105]. Lipopolysaccharide(LPS)-treated human peripheral blood monocyte (PBM) lysate (35ug protein in RIPA buffer) was resolved on a 15% SDS PAGE gel and blots probed with the rabbit IgG-chimeric version of 3F12.2D3 [NBP2-81105] at 2 ug/ml before detection using an anti-rabbit secondary antibody. Protein was detected by chemiluminescence. The expected unmodified running size for Lymphotoxin alpha is 22.3kDa, though the form shown here is likely glycosylated. NBP2-81105 successfully detected Lymphotoxin alpha in LPS-treated human PBM samples.![Flow Cytometry: Lymphotoxin-alpha/TNF-beta Antibody (3F12.2D3) - Chimeric - Azide and BSA Free [NBP2-81105]](https://resources.bio-techne.com/images/products/Lymphotoxin-alpha-TNF-beta-Antibody-3F12-2D3-Chimeric-Flow-Cytometry-NBP2-81105-img0001.jpg "Flow Cytometry: Lymphotoxin-alpha/TNF-beta Antibody (3F12.2D3) - Chimeric - Azide and BSA Free [NBP2-81105]")
Flow Cytometry: Lymphotoxin-alpha/TNF-beta Antibody (3F12.2D3) - Chimeric - Azide and BSA Free [NBP2-81105]
Flow Cytometry: Lymphotoxin-alpha/TNF-beta Antibody (3F12.2D3) - Chimeric [NBP2-81105] - Flow-cytometry using the Lymphotoxin-alpha/TNF-beta Antibody (3F12.2D3) [NBP2-81105]. U937 cells were fixed using 2% PFA, permeabilised using 0.5% Triton and stained with unimmunized rabbit IgG antibody (isotype control, black line) or the rabbit IgG-chimeric version of 3F12.2D3 (NBP2-81105, blue line) at a dilution of 1:100 for 1h at RT. After washing, bound antibody was detected using a goat anti-rabbit IgG AlexaFluor 488 antibody at a dilution of 1:1000 and cells analyzed using a FACSCanto flow-cytometer.Applications
Application
Recommended Usage
ELISA
Optimal dilutions of this antibody should be experimentally determined.
Flow Cytometry
Optimal dilutions of this antibody should be experimentally determined.
Surface Plasmon Resonance
Optimal dilutions of this antibody should be experimentally determined.
Western Blot
Optimal dilutions of this antibody should be experimentally determined.
Neutralization
Optimal dilutions of this antibody should be experimentally determined.
Application Notes
This chimeric rabbit antibody was made using the variable domain sequences of the original Mouse IgG2b format, for improved compatibility with existing reagents, assays and techniques. The properties of the chimieric 3F12 is equivalent to that of the original 3F12 mAb. 3F12 is able to bind to Th2 cells and additionally is can functionally block human LTalpha3 versus an isotype control (trastuzumab) - LTalpha3-induced ICAM upregulation is blocked on HUVEC (Human Umbilical Endothelial Cells) (IC50 0.04 nM) and LTalpha3-induced IL-8 is blocked in A549 cells. The antibody has also been used in ELISA studies to detect LT-alpha and to measure binding.
Please Note: Optimal dilutions of this antibody should be experimentally determined.
Formulation, Preparation, and Storage
Purification
Protein A purified
Formulation
PBS
Format
Azide and BSA Free
Preservative
0.02% Proclin 300
Concentration
1 mg/ml
Shipping
The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage
Store at 4C for up to 3 months. For longer storage, aliquot and store at -20C.
Background: Lymphotoxin-alpha/TNF-beta
Long Name
Tumor Necrosis Factor beta
Alternate Names
LTA, Lymphotoxin alpha, Lymphotoxinalpha, TNF-beta, tnfb, TNFSF1, TNFSF1B
Gene Symbol
LTA
Additional Lymphotoxin-alpha/TNF-beta Products
Product Specific Notices
This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.
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